RESUMO
Microalgae that have recently captivated interest worldwide are a great source of renewable, sustainable and economical biofuels. The extensive potential application in the renewable energy, biopharmaceutical and nutraceutical industries have made them necessary resources for green energy. Microalgae can substitute liquid fossil fuels based on cost, renewability and environmental concern. Microfluidic-based systems outperform their competitors by executing many functions, such as sorting and analysing small volumes of samples (nanolitre to picolitre) with better sensitivities. In this review, we consider the developing uses of microfluidic technology on microalgal processes such as cell sorting, cultivation, harvesting and applications in biofuels and biosensing.
Assuntos
Microalgas , Biocombustíveis , Biomassa , Combustíveis Fósseis , MicrofluídicaRESUMO
BACKGROUND: Fibrocytes express several chemokine receptors (CCR7 and CXCR4) that regulate their recruitment and trafficking into tissue-damage sites in response to specific chemokine gradients (CCL19 and CXCL12). OBJECTIVE: We investigated whether these chemoattractants and S100A9, through the receptor for advanced glycation end-products (RAGE; ie, its receptor), are involved in fibrocyte trafficking in patients with chronic obstructive asthma (COA) and during an acute exacerbation (AE) in patients without airflow obstruction (Asthma AE group). METHODS: We collected peripheral blood from 14 asthmatic patients with normal pulmonary function, 14 patients with COA, 11 patients in the Asthma AE group, and 14 healthy subjects. Isolated circulating fibrocytes were used for migration assay. Expression of CCR7, CXCR4, S100A9, and RAGE in fibrocytes was measured by using flow cytometry. CCL19 and CXCL12 expression in bronchial tissues was determined by using immunohistochemistry and RT-PCR. RESULTS: There were higher numbers of circulating fibrocytes in patients in the Asthma AE group and patients with COA. The expression of CXCL12 in bronchial tissues and CXCR4 in circulating fibrocytes was higher in the Asthma AE group and, to a lesser extent, in patients with COA. The expression of CCL19 in bronchial tissues and CCR7 in fibrocytes was higher in patients with COA. CXCL12/CXCR4 and CCL19/CCR7 enhanced fibrocyte transmigration in the Asthma AE group and in patients with COA, respectively. The upregulated expression of S100A9 and RAGE in fibrocytes of patients in the Asthma AE group and those with COA contributes to the enhanced basal migratory motility of fibrocytes. CONCLUSION: The CXCR4/CXCL12 axis contributes to chemotaxis of fibrocytes in patients in the Asthma AE group, whereas the CCR7/CCL19 axis plays an important role in patients with COA. S100A9 enhances the basal migratory motility of fibrocytes from patients in the Asthma AE group and patients with COA.
Assuntos
Asma/etiologia , Asma/metabolismo , Movimento Celular , Asma/patologia , Asma/fisiopatologia , Calgranulina B/metabolismo , Estudos de Casos e Controles , Movimento Celular/genética , Quimiocina CCL19/metabolismo , Quimiocina CXCL12/metabolismo , Doença Crônica , Progressão da Doença , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Receptores CCR7/metabolismo , Receptores CXCR4/metabolismo , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologiaRESUMO
Liver cancer represents a significant global burden in terms of cancer-related mortality, with resistance to anti-angiogenic drugs such as Sorafenib and Lenvatinib presenting a formidable challenge. Tumor angiogenesis, characterized by the formation of new blood vessels within tumors, plays a pivotal role in cancer progression and metastasis. Tumor endothelial cells, specialized endothelial cells lining tumor blood vessels, exhibit unique phenotypic and functional traits that drive aberrant vessel formation and contribute to therapy resistance. CD105, a cell-surface glycoprotein that is highly expressed on endothelial cells during angiogenesis, including tumor endothelial cells, regulates endothelial cell proliferation, migration, and vessel formation by modulating transforming growth factor-beta (TGF-ß) signaling pathways. Elevated CD105 expression on tumor endothelial cells correlates with increased angiogenic activity and poor prognosis in cancer patients. Targeting CD105 with antibodies presents a promising strategy to inhibit tumor angiogenesis and disrupt tumor vasculature, offering potential therapeutic benefits by interfering with the tumor microenvironment and inhibiting its progression. This study investigates tumor angiogenesis through a three-dimensional (3D) microfluidic co-culture system incorporating endothelial cells and hepatocellular carcinoma (HCC) cells. The primary focus is on the role of CD105 expression within the liver tumor microenvironment and its contribution to increased chemoresistance. Additionally, this research examines the influence of CD105 expression on the efficacy of tyrosine kinase inhibitors (TKIs) and its pivotal function in facilitating angiogenesis in liver tumors. The proposed microfluidic chip model investigates liver cancer cell interactions within a microfluidic chip model designed to simulate aspects of liver tumor angiogenesis.
Assuntos
Carcinoma Hepatocelular , Resistencia a Medicamentos Antineoplásicos , Dispositivos Lab-On-A-Chip , Neoplasias Hepáticas , Inibidores de Proteínas Quinases , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Inibidores de Proteínas Quinases/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Endoglina/metabolismo , Endoglina/antagonistas & inibidores , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/metabolismo , Sorafenibe/farmacologia , Linhagem Celular Tumoral , Compostos de Fenilureia , QuinolinasRESUMO
The acinus-mimicking microfluidic chip, which simulates the in vivo condition of the liver, was developed and reported in this paper. The gradient microenvironment of the liver acinus is replicated within this proposed microfluidic chip. The advantage of this acinus-mimicking chip is capable of adjusting the concentration gradient in a relatively short period of time at around 10 s. At the same instance the non-linear concentration gradient can be presented in the various zones within this microfluidic chip. The other advantage of this proposed design is in the convenience of allowing the direct injection of the cells into the chip. The environment within the chip is multi-welled and gel-free with high cell density. The multi-row pillar microstructure located at the entrance of the top and bottom flow channels is designed to be able to balance the pressure of the perfusion medium. Through this mechanism the shear stress experienced by the cultured cells can be minimized to reduce the potential damage flow from the perfusion process. The fluorescence staining and the observations of the cell morphology verify the life and death of the cells. The shear stress experienced by the cells in the various zones within the chip can be effectively mapped. The serum glutamic oxaloacetic transaminase (SGOT) collected from the supernatants was used to determine the effects of the degassing process and the shear stress of the medium flow on the cultured cells.
Assuntos
Fígado/citologia , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Aspartato Aminotransferases/sangue , Linhagem Celular , Dimetilpolisiloxanos/química , Desenho de Equipamento , Humanos , Perfusão , Pressão , Resistência ao Cisalhamento , Estresse Mecânico , Propriedades de SuperfícieRESUMO
The Klebsiella pneumoniae type 3 fimbriae are mainly composed of MrkA pilins that assemble into a helixlike filament. This study determined the biomechanical properties of the fimbriae and analyzed 11 site-directed MrkA mutants to identify domains that are critical for the properties. Escherichia coli strains expressing type 3 fimbriae with an Ala substitution at either F34, V45, C87, G189, T196, or Y197 resulted in a significant reduction in biofilm formation. The E. coli strain expressing MrkAG189A remained capable of producing a normal number of fimbriae. Although F34A, V45A, T196A, and Y197A substitutions expressed on E. coli strains produced sparse quantities of fimbriae, no fimbriae were observed on the cells expressing MrkAC87A. Further investigations of the mechanical properties of the MrkAG189A fimbriae with optical tweezers revealed that, unlike the wild-type fimbriae, the uncoiling force for MrkAG189A fimbriae was not constant. The MrkAG189A fimbriae also exhibited a lower enthalpy in the differential scanning calorimetry analysis. Together, these findings indicate that the mutant fimbriae are less stable than the wild-type. This study has demonstrated that the C-terminal ß strands of MrkA are required for the assembly and structural stability of fimbriae.
Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Fímbrias/metabolismo , Klebsiella pneumoniae/metabolismo , Proteínas de Bactérias/química , Biofilmes , Proteínas de Fímbrias/química , Estrutura Terciária de ProteínaRESUMO
This study investigated the structural and mechanical properties of Klebsiella pneumoniae type 3 fimbriae, which constitute a known virulence factor for the bacterium. Transmission electron microscopy and optical tweezers were used to understand the ability of the bacterium to survive flushes. An individual K. pneumoniae type 3 fimbria exhibited a helix-like structure with a pitch of 4.1 nm and a three-phase force-extension curve. The fimbria was first nonlinearly stretched with increasing force. Then, it started to uncoil and extended several micrometers at a fixed force of 66 ± 4 pN (n = 22). Finally, the extension of the fimbria shifted to the third phase, with a characteristic force of 102 ± 9 pN (n = 14) at the inflection point. Compared with the P fimbriae and type 1 fimbriae of uropathogenic Escherichia coli, K. pneumoniae type 3 fimbriae have a larger pitch in the helix-like structure and stronger uncoiling and characteristic forces.
Assuntos
Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , Proteínas de Fímbrias/química , Proteínas de Fímbrias/genética , Fímbrias Bacterianas/química , Fímbrias Bacterianas/genética , Mecânica , Microscopia Eletrônica de Varredura , Conformação Proteica , Escherichia coli Uropatogênica/metabolismo , Fatores de VirulênciaRESUMO
Globally, non-small cell lung cancer (NSCLC) is the leading cause of cancer deaths. Despite advancements in chemotherapy and targeted therapies, the 5-year survival rate has remained at 16% for the past forty years. Minimal residual disease (MRD) is described as the existence of either isolated tumour cells or circulating tumour cells in biological liquid of patients after removal of the primary tumour without any clinical signs of cancer. Recently, liquid biopsy has been promising as a non-invasive method of disease monitoring and treatment guidelines as an MRD marker. Liquid biopsy could be used to detect and assess earlier stages of NSCLC, post-treatment MRD, resistance to targeted therapies, immune checkpoint inhibitors (ICIs) and tumour mutational burden. MRD surveillance has been proposed as a potential marker for lung cancer relapse. Principally, biosensors provide the quantitative analysis of various materials by converting biological functions into quantifiable signals. Biosensors are usually operated to detect antibodies, enzymes, DNA, RNA, extracellular vesicles (EVs) and whole cells. Here, we present a category of biosensors based on the signal transduction method for identifying biosensor-based biomarkers in liquid biopsy specimens to monitor lung cancer treatment.
Assuntos
Técnicas Biossensoriais , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Biomarcadores Tumorais , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/terapia , Humanos , Biópsia Líquida , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/terapia , Neoplasia ResidualRESUMO
Cancer cell-immune cell hybrids and cancer immunotherapy have attracted much attention in recent years. The design of efficient cell pairing and fusion chips for hybridoma generation has been, subsequently, a subject of great interest. Here, we report a three-layered integrated Microfluidic Flip-Chip (MFC) consisting of a thin through-hole membrane sandwiched between a mirrored array of microfluidic channels and saw-tooth shaped titanium electrodes on the glass. We discuss the design and operation of MFC and show its applicability for cell fusion. The proposed device combines passive hydrodynamic phenomenon and gravitational sedimentation, which allows the transportation and trapping of homotypic and heterotypic cells in large numbers with pairing efficiencies of 75~78% and fusion efficiencies of 73%. Additionally, we also report properties of fused cells from cell biology perspectives, including combined fluorescence-labeled intracellular materials from THP1 and A549, mixed cell morphology, and cell viability. The MFC can be tuned for pairing and fusion of cells with a similar protocol for different cell types. The MFC can be easily disconnected from the test setup for further analysis.
Assuntos
Fusão Celular , Hidrodinâmica , Microfluídica , Células A549 , Fusão Celular/instrumentação , Sobrevivência Celular , Eletricidade , Humanos , Imageamento Tridimensional , Microfluídica/instrumentação , Células THP-1RESUMO
We develop light-driven optoelectronic tweezers based on the organic photoconductive material titanium oxide phthalocyanine. These tweezers function based on negative dielectrophoresis (nDEP). The dynamic manipulation of a single microparticle and cell patterning are demonstrated by using this light-driven optoelectronic DEP chip. The adaptive light patterns that drive the optoelectronic DEP onchip are designed by using Flash software to approach appropriate dynamic manipulation. This is also the first reported demonstration, to the best of our knowledge, for successfully patterning such delicate cells from human hepatocellular liver carcinoma cell line HepG2 by using any optoelectronic tweezers.
Assuntos
Separação Celular/instrumentação , Eletroforese em Microchip/métodos , Micromanipulação/instrumentação , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Desenho de Equipamento , Humanos , Luz , Neoplasias Hepáticas/patologia , Dispositivos Ópticos , Pinças Ópticas , Titânio/químicaRESUMO
We have designed and fabricated a novel chemotactic gradient Labchip for studying cell migration quantitatively. Owing to the great potential of garlic and its preparations in developing antiinflammatory drugs, the aim of the present study is to investigate the effect of garlic oil on the locomotion of a neutrophil-like cell by measuring the dynamic features of cell migration including migration direction, average migration speed, chemotactic index (CI), and motility index (MI) with the newly designed Labchip. We found that garlic oil treatment lowered the values of CI and MI and reduced the average speed of cell migration from 13 to 8 microm/min. The results indicate that garlic oil is a potential inhibitor for neutrophil-like cell migration and chemotactic responsiveness. By comparing with the effects of nocodazole and cytochalasin B, we also suggest that the antiinflammatory activity exhibited by garlic oil was mainly through inhibiting the assembly-disassembly processes of the cytoskeleton.
Assuntos
Compostos Alílicos/farmacologia , Quimiotaxia/efeitos dos fármacos , Dispositivos Lab-On-A-Chip , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Sulfetos/farmacologia , Células HL-60 , Humanos , Interleucina-8/farmacologia , Reologia/efeitos dos fármacosRESUMO
Agglutination is an antigen-antibody reaction with visible expression of aggregation of the antigens and their corresponding antibodies. Applications extend to the identification of acute bacterial infection, hemagglutination, such as blood grouping, and diagnostic immunology. Our finger-powered agglutination lab chip with external CMOS image sensing was developed to support a platform for inexpensive, rapid point-of-care (POC) testing applications related to agglutination effects. In this paper, blood grouping (ABO and Rh grouping) was utilized to demonstrate the function of our finger-powered agglutination lab chip with CMOS image sensing. Blood antibodies were preloaded into the antibody reaction chamber in the lab chip. The blood sample was pushed through the antibody reaction chamber using finger-powered pressure actuation to initiate a hemagglutination reaction to identify the blood type at the on-chip detection area using our homemade CMOS image sensing mini-system. Finger-powered actuation without the need for external electrical pumping is excellent for low-cost POC applications, but the pumping liquid volume per finger push is hard to control. In our finger-powered agglutination lab chip with CMOS image sensing, we minimized the effects of different finger push depths and achieved robust performance for the test results with different push depths. The driving sample volume per finger push is about 0.79 mm3. For different chips and different pushes, the driven sample volume per finger push was observed to vary in the range of 0.64 to 1.18 mm3. The red blood cells were separated from the plasma on-chip after the whole blood sample was finger pumped and before the red blood cells reached the antibody chamber via an embedded plasma-separation membrane. Our homemade CMOS image mini-system robustly read and identified the agglutination results on our agglutination lab chip.
Assuntos
Anticorpos/imunologia , Antígenos/imunologia , Eritrócitos/imunologia , Dispositivos Lab-On-A-Chip , Imagem Óptica , Testes Imediatos , Aglutinação , Reações Antígeno-Anticorpo , Eritrócitos/citologia , HumanosRESUMO
The measurement of growth factors released in a culture medium is considered to be an attractive non-invasive approach, apart from the embryo morphology, to identify the condition of an embryo development after fertilization in vitro (IVF), but the available embryo culture medium in the current method is only a few microlitres. This small sample volume, also of small concentration, makes difficult the application of a conventional detection method, such as an enzyme-linked immunosorbent assay (ELISA). A reliable detection of the growth factor from each embryo culture medium of such a small concentration hence remains a challenge. Here for the first time we report the results of measurement of not just one, but two, growth factors, human IL-1ß and human TNF-α, from an individual droplet of embryo culture medium with a bead-based digital microfluidic chip. The required sample volume for a single measurement is only 520â¯nL; the total duration of the on-chip process is less than 40â¯min. Using the culture media of human embryos with normal morphologic features, we found that the concentrations of TNF-α change little from day 3 to day 5-6, but the concentrations of IL-1ß for some embryos might double from day 3 to day 5-6. For other embryos even with similar normal morphologic features, some growth factors, such as IL-1ß, might exhibit different expressions during the culture period. Those growth factors could serve to distinguish the development conditions of each embryo, not merely from an observation of embryo morphology.
Assuntos
Técnicas Biossensoriais , Interleucina-1beta/isolamento & purificação , Microfluídica , Fator de Necrose Tumoral alfa/isolamento & purificação , Meios de Cultura/química , Feminino , Humanos , Interleucina-1beta/genética , Análise de Sequência com Séries de Oligonucleotídeos , Fator de Necrose Tumoral alfa/genéticaRESUMO
A novel electrolysis-based micropump using air bubbles to achieve indirect actuation is proposed and demonstrated. Compared with other electrochemical micropumps, our micropump can drive microfluids without inducing the pH value variation in the main channel and the choking/sticking phenomena of electrolytic bubbles. It is promising for biomedical applications, especially for blood transportation. Our proposed on-chip electrolysis-bubble actuator with the features of room temperature operation, low driving voltage, low power consumption and large actuation force not only can minimize the possibility of cell-damage but also may enable portable and implantable lab-on-a-chip microsystems. Utilizing our proposed hydrophobic trapeziform pattern located at the junction of the T-shaped microchannel, the micropump makes the pumped fluid in the main channel be isolated from the electrolytic bubbles. It can be used for a variety of applications without the constraints on the pumped liquid. Experimental results show that the liquid displacement and the pumping rate could be easily and accurately controlled via the signal of a two-phase peristaltic sequence and the periodic generation of electrolytic bubbles. With an applied voltage of 2.5 V, the maximum pumping rate for DI water and whole blood were 121 nl min(-1) and 88 nl min(-1), respectively, with a channel cross section of 100 x 50 microm. Maximum back-pressure of 16 kPa and 11 kPa for DI water and whole blood, respectively, were achieved in our present prototype chips.
Assuntos
Análise Química do Sangue/instrumentação , Eletrólise/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Dimetilpolisiloxanos/química , Desenho de Equipamento , Humanos , Concentração de Íons de Hidrogênio , Microtecnologia , Modelos Teóricos , Adesividade Plaquetária , Polietilenoglicóis/química , Pressão , TemperaturaRESUMO
A novel ac electrokinetic microfluidic driver based on alternating current electro-osmosis flow induced by asymmetrically capacitance/chemistry-modulated microelectrode arrays has been successfully developed and demonstrated. Asymmetric capacitance modulation (ACM) is made of comb electrode arrays and parts of individual electrode surfaces are modulated/deposited with a SiO(2) dielectric layer. This proposed design can be utilized to shift the optimal operation frequency of maximum velocity to a higher frequency to minimize electrolytic bubble generation and enhance micropumping performance. The pumping velocity, described in this paper, is measured via the tracing of microbeads and is a function of applied potential, signal frequency, buffer concentration, and dielectric layer thickness. A maximum pumping velocity up to 290 microm s(-1) in 5 mM buffer solution with the applied potential of 10 Vpp is observed in our prototype device, and the estimated maximum flow rate is up to 26.1 microl h(-1). This is the first successful demonstration regarding bubble-free ac electrokinetic micropumping via such asymmetrically capacitance-modulated electrode arrays. Design, simulation, microfabrication, experimental result, and theoretical model are described in this paper to characterize and exhibit the performance of the proposed novel bubble-free ac electrokinetic microfluidic driver.
Assuntos
Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Eletroquímica/instrumentação , Eletroquímica/métodos , Eletrodos , Desenho de Equipamento , Análise de Falha de Equipamento , Cinética , Modelos Teóricos , Osmose , Propriedades de SuperfícieRESUMO
A cancer immunotherapy µ-environment LabChip, equipped with titanium oxide phthalocyanine (TiOPc)-based optoelectronic tweezers (OET) to achieve direct cell-cell contact, can be used to study the interaction between immune cells and other cells for real-time analysis of NK cells' behavior. In microfluidic devices, it is difficult to solve dead zone problems and observe dynamic cell-cell interactions. We have created a stable and static culture µ-environment which can enhance NK cell activities. In addition, OET is used to solve dead zone problems by manipulating a single cell into four-leaf-clover-shaped (FLCS) microwells made of poly(ethylene glycol) diacrylate (PEG-DA) through optofluidic maskless lithography, causing direct cell-cell contact. Our design reconstructed an in vitro human immune system for the study of dynamic immunological response. When the NK cells came into contact with the target cells in the µ-environment LabChip, we observed that the target cells showed apoptotic characteristics (i.e. cell shrinkage and blebbing within 2 h and then die within 3 h). In addition, our µ-environment LabChip demonstrated higher NK cell activity compared with conventional analysis. We have created an innovative cancer immunotherapy µ-environment LabChip to provide a stable and static µ-environment for cell-cell interaction study. Furthermore, our µ-environment LabChip showed the potential to enhance NK cell activity and to study immunological interactions between immune cells and cancer cells dynamically.
Assuntos
Imunoterapia , Dispositivos Lab-On-A-Chip , Modelos Biológicos , Pinças Ópticas , Microambiente Tumoral , Animais , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Linhagem Celular , Desenho de Equipamento , Humanos , Células K562 , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Camundongos , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/imunologiaRESUMO
Biomimetic heterogeneous patterning of hepatic and endothelial cells, which start from randomly distributed cells inside the microfluidic chamber, via the chip design of enhanced field-induced dielectrophoresis (DEP) trap is demonstrated and reported in this paper. The concentric-stellate-tip electrode array design in this chip generates radial-pattern electric fields for the DEP manipulation of the live liver cells. By constructing the geometric shape and the distribution of stellate tips, the DEP electrodes enhance the desired spatial electric-field gradients to guide and snare individual cells to form the desired biomimetic pattern. With this proposed microfluidic chip design, the original randomly distributed hepatocytes inside the microfluidic chamber can be manipulated in parallel and align into the desired pearl-chain array pattern. This radial pattern mimics the lobular morphology of real liver tissue. The endothelial cells, then, are snared into the additional pearl-chain array and settle at the space in-between the previous hepatic pearl-chain array. By this cell-lab chip, we demonstrate the in vitro reconstruction of the heterogeneous lobule-mimetic radial pattern with good cell viability after cell patterning. This work reports the rapid in-parallel patterning of the dual types of live liver cells via the enhanced DEP trap inside the microfluidic chip.
Assuntos
Fígado/fisiologia , Técnicas Analíticas Microfluídicas/métodos , Materiais Biomiméticos , Adesão Celular/fisiologia , Processos de Crescimento Celular/fisiologia , Linhagem Celular Tumoral , Sobrevivência Celular/fisiologia , Simulação por Computador , Eletrodos , Eletroforese em Microchip/instrumentação , Eletroforese em Microchip/métodos , Células Endoteliais/citologia , Células Endoteliais/fisiologia , Humanos , Fígado/citologia , Técnicas Analíticas Microfluídicas/instrumentação , Polilisina/químicaRESUMO
Here, we describe a technique to manipulate a low number of beads to achieve high washing efficiency with zero bead loss in the washing process of a digital microfluidic (DMF) immunoassay. Previously, two magnetic bead extraction methods were reported in the DMF platform: (1) single-side electrowetting method and (2) double-side electrowetting method. The first approach could provide high washing efficiency, but it required a large number of beads. The second approach could reduce the required number of beads, but it was inefficient where multiple washes were required. More importantly, bead loss during the washing process was unavoidable in both methods. Here, an improved double-side electrowetting method is proposed for bead extraction by utilizing a series of unequal electrodes. It is shown that, with proper electrode size ratio, only one wash step is required to achieve 98% washing rate without any bead loss at bead number less than 100 in a droplet. It allows using only about 25 magnetic beads in DMF immunoassay to increase the number of captured analytes on each bead effectively. In our human soluble tumor necrosis factor receptor I (sTNF-RI) model immunoassay, the experimental results show that, comparing to our previous results without using the proposed bead extraction technique, the immunoassay with low bead number significantly enhances the fluorescence signal to provide a better limit of detection (3.14 pg/ml) with smaller reagent volumes (200 nl) and shorter analysis time (<1 h). This improved bead extraction technique not only can be used in the DMF immunoassay but also has great potential to be used in any other bead-based DMF systems for different applications.
RESUMO
MEMS micro-T-switches actuated via electrochemical bubbles for cell sorting applications in a monolithic chip level are proposed and successfully demonstrated. The electrolysis-bubble actuator, which has the features of low operation temperature and high surface-tension force, is developed to actuate the micro-T-switch sorting structure in our device. The double T-structure design, the T-shape microchannel with the movable micro-T-switch structure located at the junction of the T-shape microchannel, with the electrolysis-bubble actuator makes an active-binary switch function available for cell sorting applications. The room temperature operation and the low voltage required for electrolysis actuation minimize the possibility of cell-damage that happens in the conventional high electric separation instruments, such as flow cytometry. The function of our micro-T-switch chip with a low required actuation voltage of 3.0 approximately 3.5 V is demonstrated by using human hepatoma cells in this paper. The pH-value measurements characterize the pH-value variation and distribution in the actuating chambers and the mainstream microchannels to trace the possible liver-cell injury due to the pH-value variation during electrolysis-actuation operation. The 84.1% cell viability in the sorted human hepatoma cells through our micro-T-switch sorter is observed via the fluorescence assay technique. Furthermore, 70.2% of total injected cells recover in culture after sorting and grow into colonies after micro-T-switch sorting operation. In this paper, we describe the design, microfabrication, and characterization of our micro-T-switch cell-sorting chip. We also report the cell-sorting demonstration and the cell viability results for the mammalian liver cells through our micro-T-switch cell-sorting chip.
Assuntos
Separação Celular/instrumentação , Separação Celular/métodos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Linhagem Celular , Sobrevivência Celular , Eletroquímica , Hepatócitos/citologia , Humanos , Concentração de Íons de Hidrogênio , TemperaturaRESUMO
Dendritic cells/tumor fusions have shown to elicit anti-cancer immunity in different cancer types. However, the application of these vaccines for human cancer immunotherapy are limited by the instable quality and insufficient quanity of fusion cells. We present a cell electrofusion chip fabricated using soft lithography technique, which combines the rapid and precise cell pairing microstructures and the high yield electrofusion micro-electrodes to improve the cell fusion. The design uses hydrodynamic trapping in combination with positive dielectrophoretic force (pDEP) to achieve cell fusion. The chip consists of total 960 pairs of trapping channels, which are capable of pairing and fusing both homogeneous and heterogeneous types of cells. The fused cells can be easily taken out of the chip that makes this device a distinguishable from other designs. We observe pairing efficiency of 68% with fusion efficiency of 64%.
Assuntos
Fusão Celular/métodos , Hibridomas/citologia , Imunoterapia/métodos , Dispositivos Lab-On-A-Chip , Microfluídica/métodos , Fusão Celular/instrumentação , Linhagem Celular Tumoral , Humanos , Hibridomas/imunologia , Leucemia Monocítica Aguda/patologia , Neoplasias Pulmonares/patologia , Microeletrodos , Microfluídica/instrumentaçãoRESUMO
How to release growth factors (GFs) scientifically to promote stem cell proliferation and differentiation is one of the most significant research focuses in the field of regenerative medicine. In a controlled release system, growth factors, extracellular matrices or biomaterial carriers, and sometimes stem cells together form a geometric entirety. Biomaterial carriers provide GFs with a support structure to be adhered, immobilized, encapsulated or/and protected. As a unity, the release rate and rhythm of GFs on cells are normally very delicate and precise. Up to now, the best strategy for clinical applications is the combination systems that encapsulate GFs in microspheres, particularly the nano- or micro-encapsulation techniques integrated GFs with biomaterial carriers. In this mini review, we summarize the current progress in GF delivery systems for regenerative medicine and provide an outlook on two main aspects: one is the classes of stem cells and GFs that have been used frequently in regenerative medicine, including their respective application conditions and functions; the other is the controlled GF release systems, in which various GFs are released orderly and continuously without diffusing simply and rapidly, including their respective opportunities and challenges.